Method for determining genetic predisposition to delayed epidermal turnover

Abstract

To provide means of accurately and easily determining the genetic predisposition of an individual's delayed epidermal turnover.SOLUTION: Provided is a method for determining the genetic predisposition of delayed epidermal turnover, for the mitochondrial DNA contained in a DNA-containing sample collected from a subject, which comprises: detecting one or more SNP alleles selected from SNPs at the 146th, 5147th, 6962nd, 10609th, 12406th, 12882nd, 13928th, 16249th, and 16304th bases of the human mitochondrial DNA base sequence shown in a specific base sequence; and determining that the subject has a genetic predisposition of delayed epidermal turnover when the detected allele is a risk allele.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a method for determining a genetic predisposition to delayed epidermal turnover, and more particularly to a method for determining a genetic predisposition to delayed epidermal turnover by detecting mitochondrial SNPs (mtSNPs) associated with delayed epidermal turnover, and a kit used in said method. [Background technology]

[0002] Skin is roughly divided into three layers: epidermis, dermis, and subcutaneous tissue. The epidermal tissue, which is the outermost layer of the skin, has a barrier function that retains moisture and prevents the intrusion of external stimuli and foreign substances, and plays an extremely important role for the body. The epidermal tissue is composed of multiple layers of epidermal cells at different stages of differentiation (basal layer, spinous layer, granular layer, and stratum corneum) (Non-Patent Document 1). Stem cells of epidermal cells (epidermal stem cells) are present in the basal layer, which is the lowest layer of these layers, and are thought to constantly supply new cells to the tissue by repeating proliferation and differentiation as necessary (Non-Patent Document 2). As epidermal cells differentiate, they become flattened as they move from bottom to top of the tissue, and their area increases accordingly. The process from when epidermal cells are born in the basal layer to when they become dandruff in the outermost layer and fall off is called epidermal turnover. If this epidermal turnover is delayed, it is thought to cause thinning of the epidermis, thickening of the keratin, and a decrease in barrier function. Previous research has shown that the turnover time of the epidermis correlates with the area of ​​cells present in the outermost layer of the stratum corneum (corneocyte area) (Non-Patent Document 3). In other words, it is believed that the slower the turnover rate, the larger the area of ​​keratinocytes present in the outermost layer of the epidermis. Therefore, the area of ​​keratinocytes is considered to be an index for measuring the degree of turnover delay in an individual. It has been revealed that in humans, turnover tends to be delayed with aging, and an increase in the area of ​​keratinocytes is observed (Non-Patent Document 4). It is generally known that there are individual differences in the likelihood of delayed epidermal turnover (genetic predisposition), but the establishment of a technology for accurately and simply determining this genetic predisposition has not been sufficient.

[0003] Mitochondria are one of the organelles present in cells, and have their own DNA (mitochondrial DNA; mtDNA) that is different from the nucleus. mtDNA has a total length of 16,569 base pairs, a circular double-stranded structure, and encodes 13 types of oxidative phosphorylation enzymes involved in energy production, 22 types of tRNA, and 2 types of rRNA. Unlike the nuclear genome, mtDNA exists in about 2000 copies, and is therefore susceptible to damage such as oxidation, and is prone to mutation due to its weak repair activity, etc., and therefore highly polymorphic. The relationship between mtDNA polymorphism and the risk of Alzheimer's disease, diabetes, heart disease, etc., and aging phenomena has been studied, and it has been reported that it is related to metabolic syndrome, etc. (Patent Document 1). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent No. 5360853 [Non-patent literature]

[0005] [Non-Patent Document 1] Simpson CL, Patel DM, Green KJ. Deconstructing the skin:cytoarchitectural determinants of epidermal morphogenesis. Nat Rev Mol Cell Biol 2011;12:565-80. [Non-Patent Document 2] Akamatsu H, Hasegawa S, Yamada T, Mizutani H, Nakata S, Yagami A, et al. Age-related decrease in CD271(+) cells in human skin. J Dermatol 2016;43:311-3. [Non-Patent Document 3] G. L, Grove, AM Kligman. Corneocyte Size as An Indirect Measure of Epidermal Proliferative Activity. Stratum Corneum 1983;191-195. [Non-Patent Document 4] Kawai M, Imokawa G, Mizoguchi M. Analysis of facial skin properties using keratinocyte diagnosis and stratum corneum turnover measurement. Journal of the Japanese Dermatological Association 1989;99:999-1006. Summary of the Invention [Problem to be solved by the invention]

[0006] An object of the present invention is to provide a means for accurately and simply determining an individual's genetic predisposition to delayed epidermal turnover. [Means for solving the problem]

[0007] As a result of extensive research aimed at solving the above problems, the inventors discovered a specific single nucleotide polymorphism (mtSNP) in mitochondrial DNA that is associated with a genetic predisposition to delayed epidermal turnover (the likelihood of delayed epidermal turnover occurring), and thus completed the present invention.

[0008] That is, the present invention includes the following inventions. [1] A method for determining a genetic predisposition to delayed epidermal turnover, comprising the steps of: detecting alleles of one or more of the following single nucleotide polymorphisms (SNPs) in the base sequence of human mitochondrial DNA (1) to (9) in mitochondrial DNA contained in a DNA-containing sample collected from a subject; and determining that the subject has a genetic predisposition to delayed epidermal turnover if the detected allele is a risk allele. (1) A T or C SNP at the 146th base of the base sequence shown in SEQ ID NO:1 (mt146) (2) A SNP (mt5147) of G or A at the 5147th base of the base sequence shown in SEQ ID NO:1 (3) A SNP (mt6962) of G or A at the 6962nd base of the base sequence shown in SEQ ID NO:1 (4) T or C SNP at base 10609 of the base sequence shown in SEQ ID NO:1 (mt10609) (5) SNP of G or A at the 12406th base of the base sequence shown in SEQ ID NO:1 (mt12406) (6) SNP of C or T at base 12882 of the base sequence shown in SEQ ID NO:1 (mt12882) (7) A G or C SNP at the 13928th base of the base sequence shown in SEQ ID NO:1 (mt13928) (8) T or C SNP at base 16249 of the base sequence shown in SEQ ID NO:1 (mt16249) (9) T or C SNP at the 16304th base of the base sequence shown in SEQ ID NO: 1 (mt16304) [2] A method for providing cosmetics and / or food and beverages to a subject, comprising providing the subject with cosmetics and / or food and beverages having the effect of preventing and / or improving delayed epidermal turnover according to the degree of the subject's genetic predisposition to delayed epidermal turnover based on the results determined by the method described in [1]. [3] A probe having a sequence of 10 or more bases including bases 146, 5147, 6962, 10609, 12406, 12882, 13928, 16249, and 16304 in the base sequence of human mitochondrial DNA shown in SEQ ID NO: 1, or a complementary sequence thereof, and / or A kit for determining a genetic predisposition to delayed epidermal turnover, comprising primers capable of amplifying a region including bases 146, 5147, 6962, 10609, 12406, 12882, 13928, 16249, and 16304 in the base sequence of human mitochondrial DNA shown in SEQ ID NO:1. Effect of the Invention

[0009] According to the method of the present invention, by detecting the allele of a single nucleotide polymorphism (mtSNP) on mitochondrial DNA present in a biological sample from a subject, it is possible to accurately and simply determine whether the subject has a genetic predisposition to delayed epidermal turnover (whether the subject is prone to delayed epidermal turnover). Therefore, based on the result of this determination, measures can be taken early to prevent or improve delayed epidermal turnover. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] 1. Method for determining genetic predisposition to delayed epidermal turnover The method of the present invention for determining a genetic predisposition to delayed epidermal turnover comprises the steps of detecting alleles of one or more of the following single nucleotide polymorphisms (SNPs) in the base sequence of human mitochondrial DNA (1) to (9) in mitochondrial DNA contained in a DNA-containing sample collected from a subject, and determining that the subject has a genetic predisposition to delayed epidermal turnover if the detected allele is a risk allele. (1) A T or C SNP at the 146th base of the base sequence shown in SEQ ID NO:1 (mt146) (2) A SNP (mt5147) of G or A at the 5147th base of the base sequence shown in SEQ ID NO:1 (3) A SNP (mt6962) of G or A at the 6962nd base of the base sequence shown in SEQ ID NO:1 (4) T or C SNP at base 10609 of the base sequence shown in SEQ ID NO:1 (mt10609) (5) SNP of G or A at the 12406th base of the base sequence shown in SEQ ID NO:1 (mt12406) (6) SNP of C or T at base 12882 of the base sequence shown in SEQ ID NO:1 (mt12882) (7) A G or C SNP at the 13928th base of the base sequence shown in SEQ ID NO:1 (mt13928) (8) T or C SNP at base 16249 of the base sequence shown in SEQ ID NO:1 (mt16249) (9) T or C SNP at the 16304th base of the base sequence shown in SEQ ID NO: 1 (mt16304)

[0011] The above SNPs (1) to (9) may be referred to herein as mt146, mt5147, mt6962, mt10609, mt12406, mt12882, mt13928, mt16249, and mt16304, respectively.

[0012] The base numbers of human mitochondrial DNA (hmtDNA) used in the present invention refer to numbers based on the 5'-terminal base of the base sequence of NCBI Reference Sequence (ID:NC_012920.1) (SEQ ID NO:1) being regarded as the first base.

[0013] As shown in the results of the association analysis in the Examples described below, the above-mentioned nine SNPs (mtSNPs) on human mitochondrial DNA have a p-value of less than 0.05, suggesting a statistical association with a genetic predisposition to delayed epidermal turnover. Although it is possible to determine a genetic predisposition to delayed epidermal turnover with just one of the above-mentioned SNPs, the accuracy of the determination can be improved by combining two or more of them. For example, a combination of two types is preferably a combination of mt10609 and mt146.

[0014] A single nucleotide polymorphism (SNP, hereinafter sometimes referred to as "SNP") generally refers to a state in which only one site in the nucleotide sequence of a gene differs, and the site. A polymorphism generally refers to two or more alleles that exist at a frequency of 1% or more in a population.

[0015] As used herein, "allele" refers to different bases that can be taken at a certain mtSNP site, and "risk allele" refers to an allele that is more frequent in a subject group susceptible to delayed epidermal turnover than in a subject group unlikely to suffer from delayed epidermal turnover. In addition, as used herein, "genotype" refers to the type of allele at a certain mtSNP site. For example, there are two genotypes for mt146: C type and T type.

[0016] In the determination method of the present invention, "detecting an allele of a single nucleotide polymorphism (SNP)" means identifying the type of base of the allele of the SNP, and the embodiment of "detecting an allele of a single nucleotide polymorphism (SNP)" includes detecting the allele of the SNP and identifying the genotype of the SNP.

[0017] In the determination method of the present invention, when the detected allele is a risk allele, the subject is determined to have a genetic predisposition to delayed epidermal turnover (prone to delayed epidermal turnover). Specifically, for SNPs at bases 146, 5147, 6962, 10609, 12406, 12882, 13928, 16249, and 16304 of the base sequence of human mitochondrial DNA shown in SEQ ID NO: 1, if the risk allele shown in Table 1 below is detected, the subject has a genetic predisposition to delayed epidermal turnover (prone to delayed epidermal turnover) compared to when the risk allele is not detected. For example, it can be determined that there is a high possibility that delayed epidermal turnover will occur in an inappropriate manner for age, and that the possibility of delayed epidermal turnover in the future is relatively high, and it can be determined that the subject has a high degree of genetic predisposition to delayed epidermal turnover (prone to delayed epidermal turnover).

[0018] For example, in the case of mt146, the T genotype indicates that epidermal turnover delay is more likely to occur than the C genotype.

[0019] [Table 1]

[0020] In the method of the present invention, the race of the subject is not particularly limited, but is preferably East Asian, more preferably Japanese, where East Asian refers to a person originating from any of Japan, Korea, China, Taiwan, and Mongolia.

[0021] The DNA-containing sample used in the determination method of the present invention is not particularly limited as long as it is a biological sample containing DNA collected from a subject. The DNA contained in the DNA-containing sample may contain mitochondrial DNA. For example, any body fluid, secretion, tissue, cell, culture of tissue or cell, etc. from which mitochondrial DNA can be collected can be used as the DNA-containing sample. Specifically, the subject's saliva, blood, urine, sputum, pharyngeal swab, nasal swab, oral (inner cheek) mucosa swab, lacrimal gland secretion, sweat, hair, nails, skin, mucous membrane, tape strip peeled off after skin attachment, etc. can be mentioned, but saliva is preferred from the viewpoint of ease and low invasiveness. The sample can be collected according to the method used in general clinical tests and prepared using known extraction and purification methods. In this case, a commercially available genomic DNA extraction kit can be used.

[0022] The method for detecting SNPs and determining the type of SNP (allele) (SNP typing) is not particularly limited, and can be performed by known methods, such as a method in which an allele-specific primer (and probe) is used to amplify the allele by PCR or the like, and the polymorphism of the amplified product is detected by fluorescence or luminescence. For example, these include the PCR-RFLP (restriction fragment length polymerase) method, the PCR-SSCP (single-strand conformation polymerase) method, the PCR-SSO (sequence specific oligonucleotide) method, the direct sequencing method, the ASO (allele specific oligonucleotide) hybridization method, the ASP-PCR (allele specific primer-PCR) method, the SNAPSHOT method, the ARMS (amplification refraction mutation system) method, the TAQMAN PCR method, the invader method, the MALDI-TOF / MS method, the RNASE A cleavage method, the DOL (dye-labelled oligonucleotide ligation) method, and the TDI (template-directed dye-terminator incorporation). All of the above methods are well known to those skilled in the art, and reagents and kits for determining the type of SNP are commercially available. For example, TAQMAN SNP GENOTYPING ASSAYS (manufactured by Thermo Fisher Scientific) can be used.

[0023] The results obtained by the above-mentioned determination method are useful indicators for selecting cosmetics and / or foods and beverages having the effect of preventing and / or improving delayed epidermal turnover for a subject, and for example, measures to prevent delayed epidermal turnover can be recommended to a subject determined to have a genetic predisposition to delayed epidermal turnover (prone to delayed epidermal turnover), regardless of age. Thus, according to another aspect of the present invention, there is also provided a method for providing a cosmetic and / or food and beverage to a subject, the method comprising providing the subject with a cosmetic and / or food and beverage having the effect of preventing and / or improving delayed epidermal turnover according to the degree of the subject's genetic predisposition to delayed epidermal turnover (prone to delayed epidermal turnover) based on the results obtained by the above-mentioned determination method.

[0024] 2. Kit for determining genetic predisposition to delayed epidermal turnover In the above-mentioned methods for detecting and typing SNPs, probes and primers appropriate for each method are used. Such probes and primers are also included in the scope of the present invention and can be provided as a kit.

[0025] The probe may include a probe that contains the above mtSNP site and can distinguish the type of base at the mtSNP site based on the presence or absence of hybridization. Specifically, the probe may include a probe having a sequence of at least 10 consecutive bases, preferably 15 consecutive bases or more, including bases 146, 5147, 6962, 10609, 12406, 12882, 13928, 16249, and 16304 of the base sequence of human mitochondrial DNA shown in SEQ ID NO: 1, or a complementary sequence thereof. The length of the probe is preferably 15 to 40 bases, more preferably 20 to 35 bases. The probe may be labeled with an appropriate labeling substance, and examples of the labeling substance include enzymes (peroxidase, β-galactosidase, alkaline phosphatase, etc.), fluorescent substances (FITC, RITC, Cy3, Cy5, etc.), luminescent substances (luminol, luminol derivatives, luciferin, lucigenin, etc.), radioisotopes ( 3 H, 14 C. 32 P, 125 I,131 I, etc.), biotin, digoxigenin, a polypeptide containing a tag sequence, etc. Alternatively, a quencher (quenching substance) that absorbs the fluorescent energy emitted by the fluorescent substance may be further bound to the vicinity of the fluorescent substance.

[0026] The probes may also be fixed to a solid phase (DNA array). A DNA array hybridizes sample DNA to a number of probes arranged on the same plane, and by scanning the plane, it is possible to simultaneously detect hybridization to each probe. Thus, a DNA array is useful for simultaneously analyzing a number of mtSNP sites. Oligonucleotides that serve as probes to be mounted on the array are usually synthesized in situ. For example, in situ synthesis methods of oligonucleotides using a lithography method (Thermo Fisher Scientific), an inkjet method (Agilent), a bead array method (Illumina), etc. are known.

[0027] Examples of the primer include a primer that can be used in PCR for amplifying the mtSNP site, or a primer that can be used for sequence analysis (sequencing) of the mtSNP site. Specifically, examples include a primer that can amplify or sequence a region including bases 146, 5147, 6962, 10609, 12406, 12882, 13928, 16249, and 16304 of the base sequence of human mitochondrial DNA shown in SEQ ID NO: 1. The primer that can be used in PCR for amplifying the mtSNP site may be an oligonucleotide that can initiate complementary strand synthesis toward the mtSNP site using the DNA of the region including the mtSNP site as a template, and the length of such a primer is preferably 10 to 30 bases, more preferably 15 to 25 bases. The primer can be set at a position upstream or downstream of the mtSNP site in the base sequence shown in SEQ ID NO: 1.

[0028] Those skilled in the art can design probes and primers according to the analytical method based on the base sequence information of the surrounding DNA region including the mtSNP site. Furthermore, oligonucleotides to be used as probes and primers can be synthesized by a method known in the art as a method for synthesizing oligonucleotides, such as the phosphoramidite method and the H-phosphonate method, using a commonly used automatic DNA synthesizer.

[0029] The kit of the present invention may contain at least the above-mentioned probe and oligonucleotides used as primers. The kit may also contain, as necessary, a DNA extraction reagent, a PCR buffer solution, a PCR reagent such as DNA polymerase, a detection reagent such as a staining agent or an electrophoresis gel, an immobilization carrier, a labeling substance, a substrate compound used to detect the label, positive and negative standard samples, and an instruction manual that describes how to use the kit. The reagents in the kit may be in the form of a solution or a lyophilized product. EXAMPLES

[0030] The present invention will be described in more detail below with reference to examples, although the present invention is not limited to these.

[0031] (Example 1) Identification of mtSNPs associated with genetic predisposition to delayed epidermal turnover (ease of delayed epidermal turnover) In order to conduct this study, the consent form, genotype determination, etc. were approved by our company's ethical review committee. Samples were collected from subjects who had given their consent by signing the consent form, and genotype determination and association analysis with phenotype were performed.

[0032] (1)DNA sample Female employees of the company were recruited to provide DNA samples. Saliva samples were collected from 363 women who responded to the recruitment. DNA (including genomic DNA and mitochondrial DNA) was extracted from the collected saliva using the Maxwell RSC Stabilized Saliva DNA Kit (Promega Corporation) to obtain DNA samples.

[0033] (2) Amplification of human mitochondrial DNA The entire length of human mitochondrial DNA (16,569 base pairs) was amplified by long-range PCR from the obtained DNA sample. Specifically, the DNA was amplified as two fragments including overlap using the following primer sets 1 and 2. PCR was performed using SimpliAmp (Applied Biosystems) and TaKaRa LA Taq (Takara). The conditions were 94°C for 1 minute, followed by 30 cycles of 94°C for 30 seconds, 54°C for 15 seconds, and 68°C for 11 minutes, followed by reaction at 72°C for 10 minutes.

[0034] [Primer set 1] Amplification product 9064 base pairs Forward primer: 5'-AAAGCACATACCAAGGCCAC-3' (SEQ ID NO: 2) Reverse primer: 5'-TTGGCTCTCCTTGCAAAGTT-3' (SEQ ID NO: 3)

[0035] [Primer set 2] Amplification product 11170 base pairs Forward primer: 5'-TATCCGCCATCCCATACATT-3' (SEQ ID NO: 4) Reverse primer: 5'-AATGTTGAGCCGTAGATGCC-3' (SEQ ID NO: 5)

[0036] (3) Library construction Next, a library was prepared from the PCR products prepared in (2) using the Nextera XT DNA Sample Prep Kit (Illumina) and the Nextera XT Index Kit v2 Set A (Illumina). The experimental method followed the protocol of each kit.

[0037] (4) Sequence Using the library prepared in (3), the whole mitochondrial sequences of 363 subjects were analyzed using the next-generation sequencer MiSeq (Illumina). For sequencing, MiSeq Regent Kit v2 (300 cycle, Illumina) was used.

[0038] (5) Identification of mtSNPs The mtSNPs were identified by comparing the mitochondrial DNA sequence of each subject with a reference sequence (NC_012920.1; SEQ ID NO: 1). The analysis was performed using CLC Genomics Workbench (CLC bio).

[0039] (6) Phenotypic data (keratinocyte area) The keratinocyte area of ​​the subjects was measured as an index of delayed epidermal turnover. Specifically, keratinocytes were collected from the inner upper arm of the 363 subjects by tape stripping, stained by Grimelius staining, and then the cell area was measured. Grimelius staining was performed by staining the tape with keratinocytes attached with 0.03% silver nitrate solution (37°C for 3 hours), reducing solution (37°C for 3 hours), and 2% sodium thiosulfate solution (room temperature for 3 minutes), washing with running water for 5 minutes, and drying. The above-mentioned reagents were manufactured by Muto Chemical Co., Ltd. Next, 20 keratinocytes per subject were photographed using a microscope (DMI 6000B manufactured by Leica), and the area of ​​each cell was measured using image processing software ImageJ. The average value of the keratinocyte area for each subject was then calculated as the representative value for that subject.

[0040] (7) Related Analysis The association between genotype data and keratinocyte area data was evaluated using R 3.6.3 (www.r-project.org). To remove the association between phenotype data and age, a regression analysis was performed with phenotype data as the objective variable and age as the explanatory variable, and the residuals were calculated. Then, for each mtSNP, a Welch's t-test was performed on the difference in the mean residuals between the two groups divided by the presence or absence of mutation, and mtSNPs with a calculated p-value of less than 0.05 were extracted as having an association with the phenotype.

[0041] (8) Results From the above analysis, nine mtSNPs were identified that are associated with a genetic predisposition to delayed epidermal turnover (increased keratinocyte area) and can determine an individual's susceptibility to delayed epidermal turnover (Table 2). The identified mtSNPs, their positions on human mitochondrial DNA, allele and risk allele bases, and genotype p-values ​​are shown.

[0042] [Table 2]

[0043] (Example 2) Multivariate analysis of mtSNPs associated with genetic predisposition to delayed epidermal turnover Regarding the mtSNPs identified in Example 1, a combination of mtSNPs that can improve the accuracy of determining the genetic predisposition to delayed epidermal turnover was examined. Specifically, in order to remove the association between phenotype data and age, a regression analysis was performed with phenotype data as the objective variable and age as the explanatory variable, and the residual was calculated. Next, regarding the selection order of mtSNPs, a comparison was made between the case where variable selection was performed by the stepwise method (Table 3-1) and the case where variables were simply selected in ascending order of p-value of the Welch's t-test (Table 3-2) in a multiple regression analysis with the residual as the objective variable and a variable with 1 as the case where there is a mutation in the genotype data of the SNP and 0 as the explanatory variable.

[0044] [Table 3-1]

[0045] [Table 3-2]

[0046] As shown in Tables 3-1 and 3-2, the combination of mt10609 and mt146, selected as the optimal variables by the stepwise method, had a higher coefficient of determination R2 (and adjusted coefficient of determination: corrected R2), which is an index of the accuracy of the multiple regression equation, than the combination of mt10609 and mt6962, selected in ascending order of p-values ​​of the Welch t-test. In addition, the Akaike's Information Criterion (AIC) was smaller for the combination of mt10609 and mt146 than for the combination of mt10609 and mt6962, indicating a higher degree of fit of the multiple regression model. Therefore, when making a judgment by adding other mtSNPs to mt10609, it is possible to further improve the judgment accuracy by combining it with mt146. [Industrial Applicability]

[0047] The method of the present invention makes it possible to accurately and simply determine whether a sample donor has a genetic predisposition to delayed epidermal turnover, and whether or not there is a possibility and degree of delayed epidermal turnover in the future. Therefore, based on the determination result, it becomes possible to provide the sample donor with cosmetics and supplements customized for preventing or improving delayed epidermal turnover, and to provide counseling and advice regarding care methods for preventing or improving delayed epidermal turnover.

Claims

1. A method for determining a genetic predisposition to delayed epidermal turnover, comprising the steps of: detecting an allele of one or more of the following single nucleotide polymorphisms (SNPs) in a base sequence of human mitochondrial DNA (1) to (9) in mitochondrial DNA contained in a DNA-containing sample collected from a subject; and determining that the subject has a genetic predisposition to delayed epidermal turnover if the detected allele is a risk allele. (1) A T or C SNP at the 146th base of the base sequence shown in SEQ ID NO:1 (mt146, risk allele is T) (2) SNP of G or A at the 5147th base of the base sequence shown in SEQ ID NO: 1 (mt5147, risk allele is A) (3) SNP of G or A at the 6962nd base of the base sequence shown in SEQ ID NO: 1 (mt6962, risk allele is A) (4) T or C SNP at the 10609th base of the base sequence shown in SEQ ID NO: 1 (mt10609, risk allele is C) (5) SNP of G or A at the 12406th base of the base sequence shown in SEQ ID NO: 1 (mt12406, risk allele is A) (6) SNP of C or T at the 12882nd base of the base sequence shown in SEQ ID NO: 1 (mt12882, risk allele is T) (7) G or C SNP at the 13928th base of the base sequence shown in SEQ ID NO: 1 (mt13928, risk allele is C) (8) T or C SNP at the 16249th base of the base sequence shown in SEQ ID NO: 1 (mt16249, risk allele is C) (9) T or C SNP at the 16304th base of the base sequence shown in SEQ ID NO: 1 (mt16304, risk allele is C)

2. A method for providing cosmetics and / or food and beverages to a subject, comprising providing the subject with cosmetics and / or food and beverages having the effect of preventing and / or improving delayed epidermal turnover according to the degree of the subject's genetic predisposition to delayed epidermal turnover based on the results determined by the method described in claim 1.

3. A probe having a sequence of 15 to 40 consecutive bases including bases 146, 5147, 6962, 10609, 12406, 12882, 13928, 16249, and 16304 in the base sequence of human mitochondrial DNA shown in SEQ ID NO: 1, or a complementary sequence thereto, and / or A kit for determining a genetic predisposition to delayed epidermal turnover, comprising primers capable of amplifying a region including bases 146, 5147, 6962, 10609, 12406, 12882, 13928, 16249, and 16304 in the base sequence of human mitochondrial DNA shown in SEQ ID NO:1.

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